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BioAssay: AID 370

Cytotoxicity assay human pulmonary artery cells

This assay provides a robust method for measuring cytotoxicity in a readily automated 384-well format. Cell viability is determined using the CellTiter Glo reagent (Promega), which gives a luminescent readout of cellular ATP levels. ..more
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 Tested Compounds
 Tested Compounds
All(1)
 
 
Active(1)
 
 
 Tested Substances
 Tested Substances
All(1)
 
 
Active(1)
 
 
 Related BioAssays
 Related BioAssays
AID: 370
Data Source: PCMD (CHPA)
BioAssay Type: Primary, Primary Screening, Single Concentration Activity Observed
Depositor Category: NIH Molecular Libraries Screening Center Network
BioAssay Version:
Deposit Date: 2006-02-27
Modify Date: 2007-07-19

Data Table ( Complete ):           Active    All
BioActive Compound: 1
Description:
This assay provides a robust method for measuring cytotoxicity in a readily automated 384-well format. Cell viability is determined using the CellTiter Glo reagent (Promega), which gives a luminescent readout of cellular ATP levels.

Using this assay we have determined the cytotoxicity of doxorubicin against human pulmonary artery cells (HPAECs), which are a target of the aerosol delivery of therapeutic agents. Doxorubicin was provided spiked in 30 random locations in a 384-plate provided from Discovery Partners International (DPI) through the MLSCN.
Protocol
(1) Seed HPAECs in opaque clear-bottom 384-well plates (Corning) at 1250 cells/well in 20 uL media.
(2) After 24 h at 37 oC, add doxorubin in 20 uL media to give a final concentration of 2 uM. (Final DMSO concentration of 0.4 percent).
(3) Seal with gas-permeable membrane, and incubate for 72 h at 37 oC.
(4) Add 40 uL of CellTiter Glo and allow mixture to stand for at least 30 min.
(5) Read luminescence on Envision.

Data analysis:

A template written in ActivityBase (IDBS) automatically calculates percent inhibition from raw luminescence data using within-plate controls (no compound) and blanks (no cells).
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Percent inhibitionPercent inhibition at 2 uM compound. Defined as cell viability relative to viability of control cells cultured in the absence of compound. Cell viability determined by measurement of level of cellular ATP using CellTiter Glo luminescent ATP detection kit.Float%
2Mean percent inhibition: plate 1Average of percent inhibition from 30 wells containing doxorubin.Float%
3SD percent inhibition: plate 1Standard deviation of percent inhibition from 30 wells containing doxorubin.Float
4Number of replicatesInteger
5Mean percent inhibition: plate 2Average of percent inhibition from 30 wells containing doxorubin.Float%
6SD percent inhibition: plate 2Standard deviation of percent inhibition from 30 wells containing doxorubin.Float
7Number of replicatesInteger
8Mean percent inhibition control wells: plate 1Average of percent inhibition from 290 wells containing DMSO diluted to 0.4 percent but no test compound.Float%
9SD percent inhibition control wells: plate 1Standard deviation of percent inhibition from 290 wells containing DMSO diluted to 0.4 percent but no test compound.Float
10Number of replicatesInteger
11Mean percent inhibition control wells: plate 2Average of percent inhibition from 290 wells containing DMSO diluted to 0.4 percent but no test compound.Float%
12SD percent inhibition control wells: plate 2Standard deviation of percent inhibition from 290 wells containing DMSO diluted to 0.4 percent but no test compound.Float
13Number of replicatesInteger

Data Table (Concise)
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